Introduction
Elabscience® SDS-PAGE Gel
Assay kit is the classic SDS-PAGE gel preparation
kit which contains all kinds of reagents needed to prepare the SDS-PAGE gel.
Users only need apparatus and ddH2O to complete the gel preparation.
The Gel Mix in this kit is the mixture
of the gel buffer such as SDS and Tris-HCl and so on, which simplify the
procedure of gel preparation. There are Tris-HCl pH 8.8 and SDS in the Separating
Gel Mix, Tris-HCl pH 6.8 and SDS are in the Stacking Gel Mix.
Compenonts
|
Cat.
|
Products
|
10 Assays
|
25 Assays
|
50 Assays
|
Storage
|
|
E-IR-R305A
|
30% Acr-Bis (29:1)
|
45 mL
|
100 mL
|
100 mLx2
|
2~8°C
|
|
E-IR-R305B
|
Separating Gel Mix
|
22 mL
|
55 mL
|
55 mL x2
|
RT
|
|
E-IR-R305C
|
Stacking Gel Mix
|
6 mL
|
12 mL
|
22 mL
|
RT
|
|
E-IR-R305D
|
APS
|
170 mg
|
170 mg x 2
|
700 mg
|
RT
|
|
E-IR-R305E
|
TEMED
|
100 μL
|
250 μL
|
500 μL
|
2~8°C
|
|
Manual
|
One Copy
|
Instructions
According to the different molecular
weight of the target protein, different concentration of separating gel should
be prepared.
1. Comparison of protein separation
linear range is shown in the manual.
2. Sample volume of each component
corresponding to different stacking gel volumes is shown in the manual.
3. Sample volume of each component
corresponding to different separation gel volumes is shown in the manual.
Procedure
1. Preparation of 10% APS: add distilled water or DI water (add
10μL of water to 1mg APS) to APS [E-IR-R305D] to prepare a 10% APS,
For
example: add 1.7mL distilled water to170mg APS, the mixture is 10%
APS. 10% APS should be split into small tubes and stored at -20°C.
2. Choose the clean gel mold and complete the assembly.
2. Refer to in the manual, prepare the separating gel of the required concentration and volume. Add appropriate amount of ddH2O,
30% Acr-Bis (29:1)[ E-IR-R305A] and Separating Gel Mix [E-IR-R305B] to a clean beaker, mix it.
3. Add appropriate amount of 10% APS and TEMED, mix gently and avoid bubbles.
4. Add the mixture gel to the assembled gel mold immediately. Then add 1-2 mL ddH2O or absolute alcohol on the separating gel to
flatten the separating gel level.
5. Keep at RT for 30-60 min until the separating gel solidified completely.
6. Pour out the ddH2O or absolute ethanol, absorb the residual liquid with absorbent paper.
7. Refer to in the manual, prepare the stacking gel of the required volume. Add appropriate amount of ddH2O, 30% Acr-Bis (29:1)
[ E-IR-R305A] and stacking Gel Mix [E-IR-R305C] to a clean beaker, mix it.
8. Add appropriate amount of 10% APS and TEMED, mix gently and avoid bubbles.
9. Add the mixture gel upon to the separating gel immediately. Insert comb teeth carefully, avoid bubbles.
10. Keep at RT for 30-40 min until the stacking gel solidified completely.
11. Pull out the comb teeth carefully. Follow the SDS-PAGE experiment.
Storage
Store separately.
30% Acr-Bis (29:1) [E-IR-R305A] and TEMED[E-IR-R305E] should be stored at 2~8°C in dark.
10% APS should be split
into small tubes and stored at -20°C for 6
months, If
store at 2~8°C, please do not use it after one week.
Other reagents should be stored at RT.
Cautions
1. For maximal assay performance, this reagent should be used within 6 months.
Avoid freeze / thaw cycles.
2. TEMED
is volatile. Please tighten the cap after use. Due to the different
temperatures, the rate of gel solidification will be different. The amount of
TEMED can be adjusted appropriately to adjust the rate of gelation at different
temperatures.
3. Avoid
bubbles to avoid affecting the experimental results.
4. Add
the ddH2O or absolute alcohol on the separating gel gently to avoid uneven line
pressing.
5. This
kit is for research use only. For your safety and health, please wear lab
clothes and gloves. Instructions should be followed strictly, changes of
operation may result in unreliable results.
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